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目的观察弓形虫慢性感染对大鼠海马神经细胞周期、细胞凋亡及半胱氨酸蛋白酶-3(caspase-3)和细胞色素C(Cytc)表达的影响。方法将20只成年雄性SD大鼠随机分成2组:弓形虫感染组和正常对照组。弓形虫感染组每只大鼠腹腔注射纯化弓形虫速殖子2×107/ml悬液2ml;正常对照组大鼠每只腹腔注射2ml灭菌生理盐水。感染10周后应用流式细胞术检测大鼠海马神经细胞周期和细胞凋亡,罗丹明123染色检测其线粒体膜电位;应用免疫组化法检测海马caspase-3和Cytc蛋白表达。结果弓形虫感染10周后,大鼠海马神经细胞凋亡率为(49.3±13.6)%,与正常对照组(40.1±11.2)%相比较,差异有统计学意义(P<0.05),G0/G1期细胞百分率增高(P<0.05),出现G0/G1期阻滞,S期细胞减少(P<0.05);感染组线粒体膜电位为(22.1±6.8)%,比对照组的(25.4±9.4)%低(P<0.05);caspase-3和Cytc蛋白表达则均显著增高(P<0.05)。结论弓形虫慢性感染可能导致大鼠海马神经细胞凋亡,进而引起一定的认知功能障碍。
Objective To observe the effects of chronic infection with Toxoplasma gondii on the cell cycle, apoptosis and the expression of caspase-3 and Cytc in hippocampus of rats. Methods Twenty adult male Sprague Dawley rats were randomly divided into two groups: Toxoplasma gondii infection group and normal control group. Toxoplasma infection group, each mouse was injected intraperitoneally with purified Toxoplasma gondii 2 × 107 / ml suspension 2ml; normal control group rats were injected intraperitoneally 2ml sterilized saline. Ten weeks after infection, the cell cycle and apoptosis of hippocampal neurons were detected by flow cytometry. The mitochondrial membrane potential was detected by rhodamine 123 staining. The expressions of caspase-3 and Cytc in hippocampus were detected by immunohistochemistry. Results After 10 weeks of infection with Toxoplasma gondii, the apoptosis rate of hippocampal neurons in rats was (49.3 ± 13.6)%, which was significantly higher than that in control group (40.1 ± 11.2)% (P <0.05) (P <0.05). The arrest of G0 / G1 phase and the decrease of S phase cells in the control group (P <0.05). The mitochondrial membrane potential in the infected group was (22.1 ± 6.8)%, )% (P <0.05), while the expressions of caspase-3 and Cytc were significantly increased (P <0.05). Conclusion Chronic infection of Toxoplasma gondii may result in apoptosis of hippocampal neurons in rats, which may lead to certain cognitive dysfunction.